Three novel β-propeller mutations causing Glanzmann thrombasthenia result in production of normally stable pro-αIIb, but variably impaired progression of pro-αIIbβ3 from endoplasmic reticulum to Golgi

E. J.R. Nelson, J. Li, W. B. Mitchell, M. Chandy, A. Srivastava, B. S. Coller

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23 Citations (Scopus)

Abstract

Background: Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by lack of platelet aggregation in response to most physiological agonists and caused by either a lack or dysfunction of the platelet integrin αIIbβ3 (glycoprotein IIb/IIIa). Objectives: To determine the molecular basis of GT and characterize the mutations by in vitro expression studies. Patients: We studied three unrelated patients from southern India whose diagnosis was consistent with GT. Results: Immunoprecipitation of the cell lysates and immunoblotting showed no detectable mature αIIb in the G128S mutant, in contrast to 6% and 33% of the normal amount of mature αIIb in the S287L and G357S mutants, respectively. Pulse-chase analysis demonstrated pro-αIIb in the mutants comparable with the normal pro-αIIb, but no conversion to mature αIIb in the G128S mutant, and only trace conversion to mature αIIb in the S287L and G357S mutants. The disappearance of pro-αIIb in the three mutants was similar to that in cells expressing normal αIIbβ3 or αIIb only. All three mutants demonstrated pro-αIIbβ3 complexes and co-localized with an ER marker by immunofluorescence. The G128S mutant showed no co-localization with a Golgi marker, and the other two mutants showed minimal and moderate co-localization with the Golgi marker. Conclusions: These three β-propeller mutations do not affect the production of pro- αIIb, its ability to complex with β3, or its stability, but do cause variable defects in transport of pro- αIIbβ3 complexes from the endoplasmic reticulum to the Golgi.

Original languageEnglish (US)
Pages (from-to)2773-2783
Number of pages11
JournalJournal of Thrombosis and Haemostasis
Volume3
Issue number12
DOIs
StatePublished - Dec 1 2005
Externally publishedYes

Fingerprint

Thrombasthenia
Endoplasmic Reticulum
Mutation
Platelet Glycoprotein GPIIb-IIIa Complex
Platelet Aggregation
Immunoprecipitation
Immunoblotting
Integrins
Fluorescent Antibody Technique
India
Blood Platelets
Hemorrhage

Keywords

  • αβ
  • β-propeller mutations
  • Biogenesis
  • Glanzmann thrombasthenia

ASJC Scopus subject areas

  • Hematology

Cite this

@article{f38eff602d414ab191ee9a2982152795,
title = "Three novel β-propeller mutations causing Glanzmann thrombasthenia result in production of normally stable pro-αIIb, but variably impaired progression of pro-αIIbβ3 from endoplasmic reticulum to Golgi",
abstract = "Background: Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by lack of platelet aggregation in response to most physiological agonists and caused by either a lack or dysfunction of the platelet integrin αIIbβ3 (glycoprotein IIb/IIIa). Objectives: To determine the molecular basis of GT and characterize the mutations by in vitro expression studies. Patients: We studied three unrelated patients from southern India whose diagnosis was consistent with GT. Results: Immunoprecipitation of the cell lysates and immunoblotting showed no detectable mature αIIb in the G128S mutant, in contrast to 6{\%} and 33{\%} of the normal amount of mature αIIb in the S287L and G357S mutants, respectively. Pulse-chase analysis demonstrated pro-αIIb in the mutants comparable with the normal pro-αIIb, but no conversion to mature αIIb in the G128S mutant, and only trace conversion to mature αIIb in the S287L and G357S mutants. The disappearance of pro-αIIb in the three mutants was similar to that in cells expressing normal αIIbβ3 or αIIb only. All three mutants demonstrated pro-αIIbβ3 complexes and co-localized with an ER marker by immunofluorescence. The G128S mutant showed no co-localization with a Golgi marker, and the other two mutants showed minimal and moderate co-localization with the Golgi marker. Conclusions: These three β-propeller mutations do not affect the production of pro- αIIb, its ability to complex with β3, or its stability, but do cause variable defects in transport of pro- αIIbβ3 complexes from the endoplasmic reticulum to the Golgi.",
keywords = "αβ, β-propeller mutations, Biogenesis, Glanzmann thrombasthenia",
author = "Nelson, {E. J.R.} and J. Li and Mitchell, {W. B.} and M. Chandy and A. Srivastava and Coller, {B. S.}",
year = "2005",
month = "12",
day = "1",
doi = "10.1111/j.1538-7836.2005.01593.x",
language = "English (US)",
volume = "3",
pages = "2773--2783",
journal = "Journal of Thrombosis and Haemostasis",
issn = "1538-7933",
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number = "12",

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TY - JOUR

T1 - Three novel β-propeller mutations causing Glanzmann thrombasthenia result in production of normally stable pro-αIIb, but variably impaired progression of pro-αIIbβ3 from endoplasmic reticulum to Golgi

AU - Nelson, E. J.R.

AU - Li, J.

AU - Mitchell, W. B.

AU - Chandy, M.

AU - Srivastava, A.

AU - Coller, B. S.

PY - 2005/12/1

Y1 - 2005/12/1

N2 - Background: Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by lack of platelet aggregation in response to most physiological agonists and caused by either a lack or dysfunction of the platelet integrin αIIbβ3 (glycoprotein IIb/IIIa). Objectives: To determine the molecular basis of GT and characterize the mutations by in vitro expression studies. Patients: We studied three unrelated patients from southern India whose diagnosis was consistent with GT. Results: Immunoprecipitation of the cell lysates and immunoblotting showed no detectable mature αIIb in the G128S mutant, in contrast to 6% and 33% of the normal amount of mature αIIb in the S287L and G357S mutants, respectively. Pulse-chase analysis demonstrated pro-αIIb in the mutants comparable with the normal pro-αIIb, but no conversion to mature αIIb in the G128S mutant, and only trace conversion to mature αIIb in the S287L and G357S mutants. The disappearance of pro-αIIb in the three mutants was similar to that in cells expressing normal αIIbβ3 or αIIb only. All three mutants demonstrated pro-αIIbβ3 complexes and co-localized with an ER marker by immunofluorescence. The G128S mutant showed no co-localization with a Golgi marker, and the other two mutants showed minimal and moderate co-localization with the Golgi marker. Conclusions: These three β-propeller mutations do not affect the production of pro- αIIb, its ability to complex with β3, or its stability, but do cause variable defects in transport of pro- αIIbβ3 complexes from the endoplasmic reticulum to the Golgi.

AB - Background: Glanzmann thrombasthenia (GT) is an autosomal recessive bleeding disorder characterized by lack of platelet aggregation in response to most physiological agonists and caused by either a lack or dysfunction of the platelet integrin αIIbβ3 (glycoprotein IIb/IIIa). Objectives: To determine the molecular basis of GT and characterize the mutations by in vitro expression studies. Patients: We studied three unrelated patients from southern India whose diagnosis was consistent with GT. Results: Immunoprecipitation of the cell lysates and immunoblotting showed no detectable mature αIIb in the G128S mutant, in contrast to 6% and 33% of the normal amount of mature αIIb in the S287L and G357S mutants, respectively. Pulse-chase analysis demonstrated pro-αIIb in the mutants comparable with the normal pro-αIIb, but no conversion to mature αIIb in the G128S mutant, and only trace conversion to mature αIIb in the S287L and G357S mutants. The disappearance of pro-αIIb in the three mutants was similar to that in cells expressing normal αIIbβ3 or αIIb only. All three mutants demonstrated pro-αIIbβ3 complexes and co-localized with an ER marker by immunofluorescence. The G128S mutant showed no co-localization with a Golgi marker, and the other two mutants showed minimal and moderate co-localization with the Golgi marker. Conclusions: These three β-propeller mutations do not affect the production of pro- αIIb, its ability to complex with β3, or its stability, but do cause variable defects in transport of pro- αIIbβ3 complexes from the endoplasmic reticulum to the Golgi.

KW - αβ

KW - β-propeller mutations

KW - Biogenesis

KW - Glanzmann thrombasthenia

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U2 - 10.1111/j.1538-7836.2005.01593.x

DO - 10.1111/j.1538-7836.2005.01593.x

M3 - Article

C2 - 16359515

AN - SCOPUS:29244462341

VL - 3

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EP - 2783

JO - Journal of Thrombosis and Haemostasis

JF - Journal of Thrombosis and Haemostasis

SN - 1538-7933

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